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Albrecht M. Koppenhöfer - One of the best experts on this subject based on the ideXlab platform.
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Role of Plant Volatiles in Host Plant Recognition by Listronotus maculicollis (Coleoptera: Curculionidae).
Journal of Chemical Ecology, 2018Co-Authors: Olga S. Kostromytska, Cesar Rodriguez-saona, Hans T. Alborn, Albrecht M. KoppenhöferAbstract:The annual bluegrass weevil (ABW), Listronotus maculicollis Kirby, is an economically important pest of short cut turfgrass. Annual bluegrass, Poa annua L., is the most preferred and suitable host for ABW oviposition, larval survival and development. We investigated the involvement of grass volatiles in ABW host plant preference under laboratory and field conditions. First, ovipositional and feeding preferences of ABW adults were studied in a sensory deprivation experiment. Clear evidence of involvement of olfaction in host recognition by ABW was demonstrated. Poa annua was preferred for oviposition over three bentgrasses, Agrostis spp., but weevils with blocked antennae did not exhibit significant preferences. ABW behavioral responses to volatiles emitted by Agrostis spp. and P. annua were examined in Y-tube olfactometer assays. Poa annua was attractive to ABW females and preferred to Agrostis spp. cultivars in Y-tube assays. Headspace volatiles emitted by P. annua and four cultivars of Agrostis stolonifera L. and two each of A. capillaris L. and A. canina L. were extracted, identified and compared. No P. annua specific volatiles were found, but Agrostis spp. tended to have larger quantities of terpenoids than P. annua. (Z)-3-hexenyl acetate, phenyl ethyl alcohol and their combination were the most attractive compounds to ABW females in laboratory Y-tube assays. The combination of these compounds as a trap bait in field experiments attracted adults during the spring migration, but was ineffective once the adults were on the short-mown turfgrass. Hence, their usefulness for monitoring weevil populations needs further investigation.
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responses of poa annua and three bentgrass species Agrostis spp to adult and larval feeding of annual bluegrass weevil listronotus maculicollis coleoptera curculionidae
Bulletin of Entomological Research, 2016Co-Authors: Olga S. Kostromytska, Albrecht M. KoppenhöferAbstract:The annual bluegrass weevil (ABW), Listronotus maculicollis Kirby, is an economically important pest of short-cut turfgrass in Eastern North America. Wide spread insecticide resistance warrants the development of alternative management strategies for this pest. ABW damage typically occurs in areas with a high percentage of annual bluegrass, Poa annua L., the preferred ABW host. Damage to bentgrasses, Agrostis spp., is much rarer and usually less severe. To aid the implementation of host plant resistance as an alternative ABW management strategy we investigated the tolerance of three bentgrass species to ABW feeding. Responses of P. annua, creeping bentgrass, Agrostis stolonifera L., colonial bentgrass, Agrostis capillaris L., and velvet bentgrass, Agrostis canina L., to adult and larval feeding were compared in greenhouse experiments. Grass responses were measured as visual damage, dry weight of the grass stems and leaves, color, density and overall grass quality. To determine possible mechanisms of grass tolerance constitutive fiber and silicon content were also determined. The three bentgrass species tolerated 2-3 times higher numbers of ABW adults and larvae than P. annua before displaying any significant quality decrease. Creeping bentgrass had the lowest damage ratings. ABW infestation caused higher plant yield reduction in P. annua (up to 42%) than in bentgrasses. Observed differences among the grass species in fiber and silicon content in the plant tissue are unlikely to play a role in the resistance of bentgrasses to ABW. Our findings clearly show that A. stolonifera is the best grass species for the implementation of host plant resistance in ABW management.
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Ovipositional preferences and larval survival of annual bluegrass weevil, Listronotus maculicollis, on Poa annua and selected bentgrasses (Agrostis spp.).
Entomologia Experimentalis et Applicata, 2014Co-Authors: Olga S. Kostromytska, Albrecht M. KoppenhöferAbstract:The annual bluegrass weevil (ABW), Listronotus maculicollis Kirby (Coleoptera: Curculionidae), is a serious and expanding pest of short-cut turfgrass on golf courses in eastern North America. Increasing problems with the development of insecticide resistance in this pest highlights the need for more sustainable management approaches. Plant resistance is one of the most promising alternative strategies. Bentgrasses are the dominant grass species on golf course fairways, tees, and putting greens in the areas affected by ABW. But Poa annua L. (Poaceae), a highly invasive weed, often constitutes a large percentage of turf stands in short-mown golf courses and is thought to be particularly susceptible to ABW. We studied resistance to ABW in four cultivars of creeping bentgrass, Agrostis stolonifera L., and two cultivars each of colonial bentgrass, Agrostis capillaris L., and velvet bentgrass, Agrostis canina L. (Poaceae), in comparison with P. annua by addressing the three major components of resistance: antixenosis (adult ovipositional and feeding preferences), antibiosis (larval survival and growth), and grass tolerance (grass damage). Our findings suggest that antixenosis/non-preference is at least partially involved in bentgrass resistance to ABW. Even though oviposition was observed in all tested grasses, females laid significantly fewer eggs in Agrostis spp. than in P. annua. Compared to P. annua, Agrostis spp. were also less suitable for larval development with lower numbers of ABW immatures recovered and larvae weighing less and being less advanced in development. Resistance levels to ABW larvae varied significantly among Agrostis spp. and cultivars. Agrostis canina was least preferred by females for oviposition and A. stolonifera was the least suitable for larval survival and development. Agrostis spp., especially A. stolonifera, were more tolerant to ABW feeding than P. annua. Our findings suggest that reduction in P. annua and replacement with Agrostis spp., especially A. stolonifera, wherever feasible should be integral to more sustainable approaches to ABW management.
Olga S. Kostromytska - One of the best experts on this subject based on the ideXlab platform.
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Role of Plant Volatiles in Host Plant Recognition by Listronotus maculicollis (Coleoptera: Curculionidae).
Journal of Chemical Ecology, 2018Co-Authors: Olga S. Kostromytska, Cesar Rodriguez-saona, Hans T. Alborn, Albrecht M. KoppenhöferAbstract:The annual bluegrass weevil (ABW), Listronotus maculicollis Kirby, is an economically important pest of short cut turfgrass. Annual bluegrass, Poa annua L., is the most preferred and suitable host for ABW oviposition, larval survival and development. We investigated the involvement of grass volatiles in ABW host plant preference under laboratory and field conditions. First, ovipositional and feeding preferences of ABW adults were studied in a sensory deprivation experiment. Clear evidence of involvement of olfaction in host recognition by ABW was demonstrated. Poa annua was preferred for oviposition over three bentgrasses, Agrostis spp., but weevils with blocked antennae did not exhibit significant preferences. ABW behavioral responses to volatiles emitted by Agrostis spp. and P. annua were examined in Y-tube olfactometer assays. Poa annua was attractive to ABW females and preferred to Agrostis spp. cultivars in Y-tube assays. Headspace volatiles emitted by P. annua and four cultivars of Agrostis stolonifera L. and two each of A. capillaris L. and A. canina L. were extracted, identified and compared. No P. annua specific volatiles were found, but Agrostis spp. tended to have larger quantities of terpenoids than P. annua. (Z)-3-hexenyl acetate, phenyl ethyl alcohol and their combination were the most attractive compounds to ABW females in laboratory Y-tube assays. The combination of these compounds as a trap bait in field experiments attracted adults during the spring migration, but was ineffective once the adults were on the short-mown turfgrass. Hence, their usefulness for monitoring weevil populations needs further investigation.
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responses of poa annua and three bentgrass species Agrostis spp to adult and larval feeding of annual bluegrass weevil listronotus maculicollis coleoptera curculionidae
Bulletin of Entomological Research, 2016Co-Authors: Olga S. Kostromytska, Albrecht M. KoppenhöferAbstract:The annual bluegrass weevil (ABW), Listronotus maculicollis Kirby, is an economically important pest of short-cut turfgrass in Eastern North America. Wide spread insecticide resistance warrants the development of alternative management strategies for this pest. ABW damage typically occurs in areas with a high percentage of annual bluegrass, Poa annua L., the preferred ABW host. Damage to bentgrasses, Agrostis spp., is much rarer and usually less severe. To aid the implementation of host plant resistance as an alternative ABW management strategy we investigated the tolerance of three bentgrass species to ABW feeding. Responses of P. annua, creeping bentgrass, Agrostis stolonifera L., colonial bentgrass, Agrostis capillaris L., and velvet bentgrass, Agrostis canina L., to adult and larval feeding were compared in greenhouse experiments. Grass responses were measured as visual damage, dry weight of the grass stems and leaves, color, density and overall grass quality. To determine possible mechanisms of grass tolerance constitutive fiber and silicon content were also determined. The three bentgrass species tolerated 2-3 times higher numbers of ABW adults and larvae than P. annua before displaying any significant quality decrease. Creeping bentgrass had the lowest damage ratings. ABW infestation caused higher plant yield reduction in P. annua (up to 42%) than in bentgrasses. Observed differences among the grass species in fiber and silicon content in the plant tissue are unlikely to play a role in the resistance of bentgrasses to ABW. Our findings clearly show that A. stolonifera is the best grass species for the implementation of host plant resistance in ABW management.
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Ovipositional preferences and larval survival of annual bluegrass weevil, Listronotus maculicollis, on Poa annua and selected bentgrasses (Agrostis spp.).
Entomologia Experimentalis et Applicata, 2014Co-Authors: Olga S. Kostromytska, Albrecht M. KoppenhöferAbstract:The annual bluegrass weevil (ABW), Listronotus maculicollis Kirby (Coleoptera: Curculionidae), is a serious and expanding pest of short-cut turfgrass on golf courses in eastern North America. Increasing problems with the development of insecticide resistance in this pest highlights the need for more sustainable management approaches. Plant resistance is one of the most promising alternative strategies. Bentgrasses are the dominant grass species on golf course fairways, tees, and putting greens in the areas affected by ABW. But Poa annua L. (Poaceae), a highly invasive weed, often constitutes a large percentage of turf stands in short-mown golf courses and is thought to be particularly susceptible to ABW. We studied resistance to ABW in four cultivars of creeping bentgrass, Agrostis stolonifera L., and two cultivars each of colonial bentgrass, Agrostis capillaris L., and velvet bentgrass, Agrostis canina L. (Poaceae), in comparison with P. annua by addressing the three major components of resistance: antixenosis (adult ovipositional and feeding preferences), antibiosis (larval survival and growth), and grass tolerance (grass damage). Our findings suggest that antixenosis/non-preference is at least partially involved in bentgrass resistance to ABW. Even though oviposition was observed in all tested grasses, females laid significantly fewer eggs in Agrostis spp. than in P. annua. Compared to P. annua, Agrostis spp. were also less suitable for larval development with lower numbers of ABW immatures recovered and larvae weighing less and being less advanced in development. Resistance levels to ABW larvae varied significantly among Agrostis spp. and cultivars. Agrostis canina was least preferred by females for oviposition and A. stolonifera was the least suitable for larval survival and development. Agrostis spp., especially A. stolonifera, were more tolerant to ABW feeding than P. annua. Our findings suggest that reduction in P. annua and replacement with Agrostis spp., especially A. stolonifera, wherever feasible should be integral to more sustainable approaches to ABW management.
Scott E. Warnke - One of the best experts on this subject based on the ideXlab platform.
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Evaluation of Population Structure within Diploid Agrostis Germplasm Based on Miniature Inverted-Repeat Transposable Elements
Crop Science, 2012Co-Authors: Keenan Amundsen, Geunhwa Jung, Rebecca Brown, Scott E. WarnkeAbstract:The bentgrasses (Agrostis spp.) are valuable species to the turfgrass industry. The bentgrasses have complex genomes resulting from interspecifi c hybridization and polyploidization events. The New England Velvet Bentgrass collection along with diploid germplasm present in the National Plant Germplasm System represent the largest collections of publicly available diploid Agrostis germplasm within the United States. In the present study, 1225 miniature inverted-repeat transposable element (MITE)-display genetic markers were used to assess the amount of genetic diversity within a collection of 181 diploid Agrostis selections. Structure v. 2.3.3 was used to determine population structure and suggests that fi ve subpopulations best explain the genetic variation present within this germplasm. An analysis of molecular variance and principal coordinate analysis also supports the subgroupings defi ned by the Structure analysis. Nine selections were found to be genetically similar to Agrostis stolonifera L. and may be related to one of its diploid progenitors. Our improved understanding of the genetic diversity among these diploid selections, resulting from this study, will help Agrostis breeders transfer important traits from these unimproved selections for cultivar improvement.
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Agrostis Species Relationships Based on trnL-trnF and atpI-atpH Intergenic Spacer Regions
HortScience, 2012Co-Authors: Keenan Amundsen, Scott E. WarnkeAbstract:The bentgrasses (Agrostis spp.) are among the most important species to the turfgrass industry. They have complex genomes resulting from polyploidization and high rates of interspecific hybridization. An understanding of species relationships would improve the efficiency of developing improved bentgrass cultivars. To elucidate the evolutionary relationships among Agrostis species, phylogenetic analyses were performed on sequences of two chloroplast-encoded intergenic spacer regions. A 298-bp region of the trnL-trnF intergenic spacer and a 451-bp region of the atpI-atpH intergenic spacer were included in the analyses. A total of 47 Agrostis accessions were included with both cultivated and unimproved material from the National Plant Germplasm System. Of these 47 Agrostis collections, there were 10 unique trnL-trnF haplotypes and eight distinct atpI-atpH haplotypes, indicating a high degree of shared sequence identity within these chloroplast intergenic regions. These findings suggest that the chloroplast genomes of A. canina and A. stolonifera are more closely related to each other than either species is to A. capillaris, incongruent with our previous understanding of genome relationships in the genus.
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Species Relationships in the Genus Agrostis Based on Flow Cytometry and MITE-Display Molecular Markers
Crop Science, 2011Co-Authors: Keenan Amundsen, Scott E. WarnkeAbstract:It is estimated that there are between 150 and 200 species of Agrostis, and interspecific hybridization is a proven method for improving cultivated Agrostis species. The pool of publicly available Agrostis germplasm, available through the National Plant Germplasm System, represents 17% of the total number of Agrostis species. The total amount of genetic diversity of this germplasm should be assessed to determine if there are novel sources of stress resistance. Agrostis species have a base haploid chromosome number of seven and can range in ploidy from diploid (2n = 2x = 14) to dodecaploid (2n = 12x = 84). In the present study, the DNA content of 305 Agrostis accessions was measured by flow cytometry and used to predict ploidy. Ploidy assignment was difficult because the DNA content measures did not delineate the accessions into clear genome size groupings. Genetic diversity among a subset of those Agrostis accessions (75), along with four Polypogon and two Apera accessions, was also studied. Genetic markers anchored to miniature inverted-repeat transposable elements (1309) were used in Structure 2.3.3 and unweighted pair-grouped method analyses. Eight distinct clusters of accessions were observed, including two distinct groups of diploid germplasm. Little genetic variation was evident among the cultivated A. stolonifera accessions based on the Structure analysis, suggesting the early formation of a genetic bottleneck among cultivated germplasm.
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Miniature Inverted‐Repeat Transposable Element Identification and Genetic Marker Development in Agrostis
Crop Science, 2011Co-Authors: Keenan Amundsen, David Rotter, Joachim Messing, Geunhwa Jung, Faith C. Belanger, Scott E. WarnkeAbstract:Creeping bentgrass (Agrostis stolonifera L.) is an important species to the turfgrass industry because of its adaptation for use in high quality turf stands such as golf course putting greens, tees, and fairways. Creeping bentgrass is a highly outcrossing allotetraploid, making genetic marker development difficult. Genetic markers anchored to miniature inverted-repeat transposable elements (MITEs) were developed in Agrostis that could be used in genetic linkage mapping, quantitative trait loci studies, or diversity analyses. The FindMITE software program identified 495 candidate M ITEs from 16,122 Agrostis DNA sequences. There was evidence of transposition in 79 of the candidate MITEs based on MITE insertional polymorphisms. Genetic markers were developed by MITE-display, a modified amplified fragment length polymorphism technique that anchors amplified fragments to MITEs. Four MITE-display primer combinations were tested on a creeping bentgrass experimental mapping population and 139 polymorphic markers were developed with a polymorphic information content of 0.33. Twenty-eight of the polymorphic genetic markers segregated normally. MITE-display genetic markers are a new class of genetic markers for studies of the Agrostis genome. These genetic markers target transposable elements and provide an easy method of identifying allelic variation between Agrostis accessions that may be used for diversity studies or genetic linkage map development.
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miniature inverted repeat transposable element identification and genetic marker development in Agrostis
Crop Science, 2011Co-Authors: Keenan Amundsen, David Rotter, Joachim Messing, Geunhwa Jung, Faith C. Belanger, Scott E. WarnkeAbstract:Creeping bentgrass (Agrostis stolonifera L.) is an important species to the turfgrass industry because of its adaptation for use in high quality turf stands such as golf course putting greens, tees, and fairways. Creeping bentgrass is a highly outcrossing allotetraploid, making genetic marker development difficult. Genetic markers anchored to miniature inverted-repeat transposable elements (MITEs) were developed in Agrostis that could be used in genetic linkage mapping, quantitative trait loci studies, or diversity analyses. The FindMITE software program identified 495 candidate M ITEs from 16,122 Agrostis DNA sequences. There was evidence of transposition in 79 of the candidate MITEs based on MITE insertional polymorphisms. Genetic markers were developed by MITE-display, a modified amplified fragment length polymorphism technique that anchors amplified fragments to MITEs. Four MITE-display primer combinations were tested on a creeping bentgrass experimental mapping population and 139 polymorphic markers were developed with a polymorphic information content of 0.33. Twenty-eight of the polymorphic genetic markers segregated normally. MITE-display genetic markers are a new class of genetic markers for studies of the Agrostis genome. These genetic markers target transposable elements and provide an easy method of identifying allelic variation between Agrostis accessions that may be used for diversity studies or genetic linkage map development.
Keenan Amundsen - One of the best experts on this subject based on the ideXlab platform.
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Evaluation of Population Structure within Diploid Agrostis Germplasm Based on Miniature Inverted-Repeat Transposable Elements
Crop Science, 2012Co-Authors: Keenan Amundsen, Geunhwa Jung, Rebecca Brown, Scott E. WarnkeAbstract:The bentgrasses (Agrostis spp.) are valuable species to the turfgrass industry. The bentgrasses have complex genomes resulting from interspecifi c hybridization and polyploidization events. The New England Velvet Bentgrass collection along with diploid germplasm present in the National Plant Germplasm System represent the largest collections of publicly available diploid Agrostis germplasm within the United States. In the present study, 1225 miniature inverted-repeat transposable element (MITE)-display genetic markers were used to assess the amount of genetic diversity within a collection of 181 diploid Agrostis selections. Structure v. 2.3.3 was used to determine population structure and suggests that fi ve subpopulations best explain the genetic variation present within this germplasm. An analysis of molecular variance and principal coordinate analysis also supports the subgroupings defi ned by the Structure analysis. Nine selections were found to be genetically similar to Agrostis stolonifera L. and may be related to one of its diploid progenitors. Our improved understanding of the genetic diversity among these diploid selections, resulting from this study, will help Agrostis breeders transfer important traits from these unimproved selections for cultivar improvement.
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Agrostis Species Relationships Based on trnL-trnF and atpI-atpH Intergenic Spacer Regions
HortScience, 2012Co-Authors: Keenan Amundsen, Scott E. WarnkeAbstract:The bentgrasses (Agrostis spp.) are among the most important species to the turfgrass industry. They have complex genomes resulting from polyploidization and high rates of interspecific hybridization. An understanding of species relationships would improve the efficiency of developing improved bentgrass cultivars. To elucidate the evolutionary relationships among Agrostis species, phylogenetic analyses were performed on sequences of two chloroplast-encoded intergenic spacer regions. A 298-bp region of the trnL-trnF intergenic spacer and a 451-bp region of the atpI-atpH intergenic spacer were included in the analyses. A total of 47 Agrostis accessions were included with both cultivated and unimproved material from the National Plant Germplasm System. Of these 47 Agrostis collections, there were 10 unique trnL-trnF haplotypes and eight distinct atpI-atpH haplotypes, indicating a high degree of shared sequence identity within these chloroplast intergenic regions. These findings suggest that the chloroplast genomes of A. canina and A. stolonifera are more closely related to each other than either species is to A. capillaris, incongruent with our previous understanding of genome relationships in the genus.
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Species Relationships in the Genus Agrostis Based on Flow Cytometry and MITE-Display Molecular Markers
Crop Science, 2011Co-Authors: Keenan Amundsen, Scott E. WarnkeAbstract:It is estimated that there are between 150 and 200 species of Agrostis, and interspecific hybridization is a proven method for improving cultivated Agrostis species. The pool of publicly available Agrostis germplasm, available through the National Plant Germplasm System, represents 17% of the total number of Agrostis species. The total amount of genetic diversity of this germplasm should be assessed to determine if there are novel sources of stress resistance. Agrostis species have a base haploid chromosome number of seven and can range in ploidy from diploid (2n = 2x = 14) to dodecaploid (2n = 12x = 84). In the present study, the DNA content of 305 Agrostis accessions was measured by flow cytometry and used to predict ploidy. Ploidy assignment was difficult because the DNA content measures did not delineate the accessions into clear genome size groupings. Genetic diversity among a subset of those Agrostis accessions (75), along with four Polypogon and two Apera accessions, was also studied. Genetic markers anchored to miniature inverted-repeat transposable elements (1309) were used in Structure 2.3.3 and unweighted pair-grouped method analyses. Eight distinct clusters of accessions were observed, including two distinct groups of diploid germplasm. Little genetic variation was evident among the cultivated A. stolonifera accessions based on the Structure analysis, suggesting the early formation of a genetic bottleneck among cultivated germplasm.
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Miniature Inverted‐Repeat Transposable Element Identification and Genetic Marker Development in Agrostis
Crop Science, 2011Co-Authors: Keenan Amundsen, David Rotter, Joachim Messing, Geunhwa Jung, Faith C. Belanger, Scott E. WarnkeAbstract:Creeping bentgrass (Agrostis stolonifera L.) is an important species to the turfgrass industry because of its adaptation for use in high quality turf stands such as golf course putting greens, tees, and fairways. Creeping bentgrass is a highly outcrossing allotetraploid, making genetic marker development difficult. Genetic markers anchored to miniature inverted-repeat transposable elements (MITEs) were developed in Agrostis that could be used in genetic linkage mapping, quantitative trait loci studies, or diversity analyses. The FindMITE software program identified 495 candidate M ITEs from 16,122 Agrostis DNA sequences. There was evidence of transposition in 79 of the candidate MITEs based on MITE insertional polymorphisms. Genetic markers were developed by MITE-display, a modified amplified fragment length polymorphism technique that anchors amplified fragments to MITEs. Four MITE-display primer combinations were tested on a creeping bentgrass experimental mapping population and 139 polymorphic markers were developed with a polymorphic information content of 0.33. Twenty-eight of the polymorphic genetic markers segregated normally. MITE-display genetic markers are a new class of genetic markers for studies of the Agrostis genome. These genetic markers target transposable elements and provide an easy method of identifying allelic variation between Agrostis accessions that may be used for diversity studies or genetic linkage map development.
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miniature inverted repeat transposable element identification and genetic marker development in Agrostis
Crop Science, 2011Co-Authors: Keenan Amundsen, David Rotter, Joachim Messing, Geunhwa Jung, Faith C. Belanger, Scott E. WarnkeAbstract:Creeping bentgrass (Agrostis stolonifera L.) is an important species to the turfgrass industry because of its adaptation for use in high quality turf stands such as golf course putting greens, tees, and fairways. Creeping bentgrass is a highly outcrossing allotetraploid, making genetic marker development difficult. Genetic markers anchored to miniature inverted-repeat transposable elements (MITEs) were developed in Agrostis that could be used in genetic linkage mapping, quantitative trait loci studies, or diversity analyses. The FindMITE software program identified 495 candidate M ITEs from 16,122 Agrostis DNA sequences. There was evidence of transposition in 79 of the candidate MITEs based on MITE insertional polymorphisms. Genetic markers were developed by MITE-display, a modified amplified fragment length polymorphism technique that anchors amplified fragments to MITEs. Four MITE-display primer combinations were tested on a creeping bentgrass experimental mapping population and 139 polymorphic markers were developed with a polymorphic information content of 0.33. Twenty-eight of the polymorphic genetic markers segregated normally. MITE-display genetic markers are a new class of genetic markers for studies of the Agrostis genome. These genetic markers target transposable elements and provide an easy method of identifying allelic variation between Agrostis accessions that may be used for diversity studies or genetic linkage map development.
Masashi Ugaki - One of the best experts on this subject based on the ideXlab platform.
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Transgenic plants of Agrostis alba obtained by electroporation-mediated direct gene transfer into protoplasts
Plant Cell Reports, 1994Co-Authors: Yoshito Asano, Masashi UgakiAbstract:A system for genetic transformation of Agrostis alba plants by electroporation-mediated DNA transfer to protoplasts is described. The npt II gene was used as a selectable marker. Selection with 20 mg/1 G418 (geneticin) yielded a total of over 50 resistant cell colonies from three independent experiments. Overall frequency of resistant colony formation was 1–3 × 10^−6 based on the number of protoplasts plated and 1–2 × 10^−5 based on the number of cell colonies recovered. Subsequent subcultures led to the development of plants with an apparently normal morphology. DNA analysis (PCR and Southern hybridization) and enzymatic analysis showed that the G418 resistant plants carried the transgene and expressed it. This is the first successful genetic transformation of an economically important temperate grass, Agrostis.
